OLIGOBIND® APC activity assay
INTENDED USE
The OLIGOBIND® APC activity assay is an enzyme-capture-assay for the quantitative measurement of active Protein C in stabilized plasma samples.
EXPLANATION OF THE TEST
Activated protein C (APC) is a multifunctional serine protease with anticoagulant, anti-inflammatory and anti-apoptotic functions. The active protein is generated from its precursor zymogen protein C (PC) on the surface of endothelial cells by an activation complex formed by thrombin and thrombomodulin (TM).(1) APC released from the endothelial cell surface down-regulates thrombin formation by cleavage of the activated cofactors V and VIII.(1,2)
A failure to generate sufficient amounts of APC is associated with a prothrombotic and a hyperinflammatory phenotype. The severity of the clinical symptoms depends on the residual APC activity. The prothrombotic phenotype is the leading symptom in milder forms of APC deficiency such as in heterozygous PC deficiency, whereas more severe forms of APC deficiency such as in homozygous PC deficiency are characterized by a thrombo-inflammatory phenotype.(2) Acquired APC dysfunction is critically involved in the pathogenesis of several thrombo-inflammatory diseases including severe sepsis.(3)
In spite of the important physiological functions of APC and its crucial role in the pathogenesis of a variety of diseases, plasma levels of APC have not been considered in diagnostic or therapeutic decision making. In combination with the APC blood collection tubes (Product No. ADG855T) that ensure the stabilization of APC-activity ex vivo, the OLIGOBIND® APC activity assay allows the direct quantification of functional active Protein C in plasma from peripheral blood.
PRINCIPLE OF THE METHOD
Stabilized plasma samples are added to microwells coated with a DNA-aptamer against APC.(4) During an incubation period, APC present in the sample will bind to the aptamer coated to the wells. Following a washing step, a fluorogenic peptide substrate for activated Protein C is added to the microwells. Measuring the change of fluorescence (360[ex]/460[em] nm) and extrapolating the value with those of a standard curve determines the level of APC in the plasma sample.
Category: Research use only
Type: Activity assay
Product Availability: Worldwide
Manufacturer: ImmBioMed GmbH & Co KG, Germany
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OLIGOBIND APC activity assay
Cat.No. ADG855
Article no.: 938250
Unit: 96 Tests
Code: ADG855
Manufacturer: ImmBioMed GmbH & Co. KG
BIBLIOGRAPHY
- Do-all receptor takes on coagulation, inflammation. Esmon CT, Nat Med 2005; 11: 475–477.
- The Protein C pathway. Esmon CT, Chest 2003; 124: 26S–32S.
- New players in the sepsis-protective activated protein C pathway. Ruf W, J Clin Invest 2010; 120: 3084–3087.
- An exosite-specific ssDNA aptamer inhibits the anticoagulant functions of activated protein C and enhances inhibition by protein C inhibitor. Müller J et al., Chem Biol 2009; 16: 442–451.
- Monitoring of plasma levels of activated protein C using a clinically applicable oligonucleotide-based enzyme capture assay. Müller J et al., J Thromb Haemost. 2012 Mar;10(3):390-398.
- Activity pattern analysis indicates increased but balanced systemic coagulation activity in response to surgical trauma. Friedrich et al., TH Open 2018; 2: e350-e356.
- In vivo thrombin generation and subsequent APC formation are increased in factor V Leiden carriers. Rühl H, Winterhagen FI, Berens C, Müller J, Oldenburg J, Pötzsch B. Blood. 2018 Mar 29;131(13):1489-1492.